JPS59164795A - Reduction method of metal complex - Google Patents

Abstract

PURPOSE:A metal complex is embedded in liposome together with a reduction assistant which has an oxidation-reduction potential more negative than the metal and is reversibly oxidative and reductive and reduction is effected at a potential more negative than the potential of the assistant whereby a metal complex used as an oxygen adsorbent is efficiently reduced. CONSTITUTION:In the reduction of a fat-soluble metal complex embedded in liposome or the hydrophobic zone in the lamella composed of mainly phospholipid by the action of outside reducing force, a fat-soluble reduction assistant, which has an redoxy potential more negative than the potential of the center metal of the complex and is reversibly oxidative and reductive, is previously embedded in the liposome or lamella, then a reducing force whose potential is more negative than that of the assistant is made to act outside to effect the reduction of the center metal of the complex which is in the oxidized state.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for reducing a metal complex embedded in the hydrophobic region of a liposome or lamella containing phospholipids as a main component.

Many metal 5-complexes are known that exhibit activity when the central metal is in a reduced state. However, when placed directly in an aqueous medium, these metal complexes quickly lose their activity and are often unable to perform their intended functions. For this reason, it is conceivable to embed a metal complex in the hydrophobic region of a phospholipid-based rhodesome or lamella and disperse it in an aqueous medium, thereby expressing a desired function even in an aqueous medium. However, even when embedded in such liposomes or lamellae, metal complexes can be oxidized, albeit gradually. In order to reduce the central metal of the oxidized complex, for example, Na2S2 is added to the aqueous medium.
It is possible to add a reducing agent such as O4, but even if such a strong reducing agent is used, it cannot reach the hydrophobic region of the liposome or lamella that embeds the metal complex because it is hydrophilic. , reduction is virtually impossible to achieve. Such problems can be solved by first embedding a metal complex having a central metal in an oxidized state in liposomes or lamellae as described above, and then reducing the central metal in an aqueous medium6.
- Occurs also in cases.

Therefore, the purpose of this invention is to provide a method for quickly and efficiently reducing in an aqueous medium a metal complex in which the central metal is embedded in an oxidized state in the hydrophobic region of a phospholipid-based liposome or lamella. It is about providing.

In order to achieve the above-mentioned object, the present invention aims at reducing the amount of fat-soluble metal complexes embedded within the hydrophobic regions of liposomes or lamellae containing phospholipids as a main component in an aqueous medium by the action of an external reducing force. In the reducing method, a liposome or lamella is pre-embedded with a fat-soluble reducing agent which has a negative redox potential compared to the redox potential of the central metal during disassembly of the metal and has reversible redox ability. Then, a reducing power corresponding to a negative redox potential of the reducing agent is applied externally to the liposome or lamella to reduce the center of the metal complex having the central metal in an oxidized state. The present invention provides a method for reducing a metal complex, which is characterized by reducing a metal.

This invention utilizes reducing power (electrons) from outside the liposome or lamella (i.e., in the aqueous medium) in an aqueous medium by embedding a metal complex or a reducing agent in a desome or lamella. By transmitting it to the metal complex via the reduction aid, it is possible to efficiently reduce the metal complex within the liposome or lamella, which has been difficult in the past. That is, in this specification, the term "supporting agent" means a substance that serves as a medium for receiving and receiving electrons involved in reduction. For this reason, the reducing aid must be lipophilic in order to be able to redox itself reversibly and to exist at least in the hydrophobic region of the liposome or lamella. Furthermore, if the external reducing power or the redox potential of the external reducing agent, reduction aid, and metal complex are respectively El, R2, and R5, then the relationship between these is E1<R2<Es.
(1) must be satisfied.

That is, the reduction aid used in this invention can be determined depending on the type of metal complex to be reduced, and the external reducing agent is also required depending on the type of metal complex to be reduced. especially,
The reduction aid preferably used in this invention is a quinone compound, and examples thereof are as follows: (1) Formula (where R1 to R4 are hydrogen, halodane, benzyl group, A benzoquinone compound represented by an alkyl group, alkenyl group, or alkynyl group having 5 or less carbon atoms.

(11) A naphthoquinone compound represented by the formula (wherein R1 to R6 are each hydrogen, halodane, benzyl group, or an alkyl group, alkenyl group, or alkynyl group having 5 or less carbon atoms).

(11) An anthraquinone compound represented by the formula (wherein R1 to R8 are each hydrogen, halodane, benzyl group, or an alkyl group, alkenyl group, or alkynyl group having 5 or less carbon atoms).

(Formula 9 (where R1 to B I2 are each hydrogen, 21
0rn, benzyl group, alkyl group, alkenyl group or alkynyl group having 5 or less carbon atoms, or alkyloxylic group having 5 or less carbon atoms? A merocyanine dye compound represented by a merocyanine group, an alkylamido group, an alkenyloxycarbunyl group, an alkenylamide group, an alkynyloxycardenyl group, or an alkynylamide group.

Formula 0 (where R1 to R7 are hydrogen, halodane,
A gallocyanine dye compound represented by a benzyl group or an alkyl group, alkenyl group, or alkynyl group having 5 or less carbon atoms, and where X is oxygen or sulfur.

A flavin dye compound represented by the formula υ (where R to R are hydrogen, halodane, benzyl group, or an alkyl group, alkenyl group, or alkynyl group having 5 or less carbon atoms).

Specific examples of such quinone compounds include duroquinone, vitamin 1, R2 and 3, anthraquinone, riboflavin, hypoxanthine, eosin Y methyl ester, flavin mononucleotide, nicotinamide adenine dinucleotide, and flavin adenine dinucleotide. These include, but are not limited to, nucleotides, vitamin B2, etc.

The reducing aid described above penetrates the lipid layer of the liposome or lamella, which will be described in detail later, and is in a superimposed association state (hereinafter referred to as stack) from the external aqueous phase of the liposome or lamella to at least the vicinity of the metal complex present in the hydrophobic region. Present within liposomes or lamellae. The amount of the reduction aid embedded in the ridesome increases, for example, as the number of carbon atoms in the substituents R1 to R12 of the quinone-based substances represented by formulas (I) to (B) increases; however, on the other hand, these substituents If the material becomes bulky, it becomes difficult to form the above-mentioned stack. The definitions of the substituents R1 to R12 in the formulas (I) to (B) are given in order to satisfactorily satisfy both of these requirements. Note that the influence of these substituents on the redox potential of the reduction aid is on the positive side when it is a methyl group, on the negative side when it is a halodane group, and on the negative side in the case of other substituents.

The metal complex to be reduced according to the present invention is fat-soluble because it is embedded in the hydrophobic regions of liposomes or lamellae containing phospholipids as a main component. When the quinone compound is used as a reduction aid, it is desirable that the metal complex has a redox potential of about -0.35 delts or more positive. This is because R2 of hypoxanthine, which has the lowest redox potential among the quinone-based 13-compounds, is -0,37&lt; Such metal complexes contain highly hydrophobic ligands such as porphyrin, chlorin, choline or derivatives thereof, N, N'-
Metal complexes with bis(salicylidene) alkylene or aralkylene diamines, 2,2'-bipyridine, 0-phenanthroline, etc., without a counterion or with halodane Cl-04-+ BF4- s P
F6-, preferably with PF6- as a counterion,
Alternatively, it is a metal complex with a ligand such that the total charge becomes O as a result of complex formation. To illustrate these, 'Fe(1
! D (tetraphenylporphine)・Br% R6 (to) (chlorophyllin)・Dr% Fe(2) (glotoporphyrin ■)・Ct% Fe(e) (N, N-bis(
salicylidene) alkylene or aralkylene diamine) ・C1, Fe0ID (2,2'-bipyridine)3
・(PF6)3, RuQID (2,2′-bipyridine)g ・(PF6)! , O@(110(2,2'-bipyridine)3 ・(PF6)!, Cu(II) (dimethylglyoxime)2 , CoQID (L-alanine)
3 etc., but is not limited to these.

14- As mentioned above, the phospholipids or lamellae that embed the reducing aid and the metal complex are formed mainly from phospholipids. Examples of phospholipids include phosphatidylinositol, sphine-2 miniphosphorus, and di(CI2-C22
Alkyl acyl)glycerophosphatidylcholine and ethanolamines include, but are not limited to, egg yolk lecithin, soybean lecithin, noharmitoylphosphatinorcholine, somyristoylphosphatidylcholine, distearoylphosphatidylcholine, phosphatidylethanolamine, and the like.

What about the reducing aids, metal complexes, and phospholipids mentioned above?
In order to form somesomes or lamellae, first, the above formula (
1) Noroform, dichloromethane, ether, alcohol, benzene, T)IF, It is uniformly dissolved in an organic solvent such as DMF, and this solution is gently concentrated under reduced pressure to form a film on the wall of the vessel. An appropriate amount of pure water or a buffered aqueous solution having a pH of about 6 to 8 is added to this and subjected to ultrasonic stirring, or the film-like material is appropriately pulverized in advance and then subjected to a crushing operation using a French pressure cell or the like. The degree of ultrasonic stirring and crushing operations depends on the concentration and the fatty chain length of the phospholipid, but it is sufficient to perform ultrasonic stirring at a power of 60 W or more for 30 minutes or more, or to crush at 20,000 ps1 or more. The liposome or lamellar aqueous dispersion obtained at this time has a hook-like appearance up to about 10% by weight, and if it exceeds this, some sedimentation occurs.

When the central metal of the metal complex used to prepare this liposome or lamellar aqueous dispersion is in an oxidized state from the beginning or becomes oxidized later, the liposome or lamellar aqueous dispersion is aerated with an inert gas. Alternatively, the central metal is quickly reduced by applying a reducing force to the dispersion after deoxidizing it by degassing under reduced pressure. Such reducing power is due to the presence of chemicals such as Na2S2 solubilized in aqueous solution.
O4a Na25O3s Na2S2O5p NaBH
4. Aruiha Crucose-6-Phosphate Toglucose Mountain 6
- a combination of phosphate dehydrodanases or NAD;
Enzymatic reducing agent group (mixed with P, ferredoxin, ferredoxin-NADP reductase, catalase, etc.)
E. Hasegawa et al., Biochem.

Biophys, Res, Commun, p
104793 (1982)), a combination of pt or Pd-carbon and hydrogen gas, etc. The most potent of these is NaBH4. During reduction, the reducing agent stack that penetrates the phospholipid layer and is exposed to the outside of the liposome or lamella, that is, to the aqueous dispersion medium, receives electrons from the reducing agent, and transfers them to the hydrophobic region of the liposome or lamella. It is thought that the central metal is reduced by passing to a nearby metal complex.

The thus obtained liposome or lamellar dispersion containing a metal complex in a low oxidation state in the phospholipid hydrophobic region has the advantage that reactions that originally dislike the intrusion of water can be carried out in an aqueous solution. - A typical example is the adsorption/desorption reaction of oxygen molecules.

For example, meso-tetra [
α, α, α, α (O-bipalamidophenyl)] The 1-alkylimidazole complex of porphinate iron (n) is dehydrated because the bimolecular oxidation reaction of the following formula (2) is prevented. A reversible oxygen molecule adsorption/desorption reaction of formula (3) has been reported in an organic solvent (J, P, Col1man
Others - J, Am, Chem, Soc, y 97
+1427 (1975)). However, L represents an imidazole-based axial ligand, and P represents a porphyrin derivative.

L-Fe(II)P+02 Sobenrei-Fe(II)P-
02(3) However, this complex does not dissolve in water at all, and when a small amount of water is poured into an oxygenated organic solvent solution, the following formula (
The oxidation reaction of 4) occurs and the capacity is immediately lost.

H” L-Fe(IT)P+02:L-Fe([)P-02→
L-FeQ[)P+ HO2° (4) 18- If the above iron complex is embedded in the hydrophobic part of phospholipid liposomes or lamellae, an apparent aqueous solution can be obtained, and the formula (
The reaction 4) is prevented and reversible oxygen adsorption/desorption becomes possible. However, even if the starting iron complex is iron ω),
During the liposome or lamella formation operation, the iron(II) state is oxidized and embedded, so it is necessary to return it to the iron(II) state by the action of an external reducing force. will be extremely useful.

Examples of metal complexes that are embedded in the hydrophobic regions of liposomes or lamellae containing phospholipids as a main component and perform reversible oxygen absorption and desorption are as follows.

(4) General formula (where each Rtri C1~CIB alkyl group, C1~
C2° ω-hydroxyalkyl group, C1-C11 ω
-Hydroxycardenylalkyl group and its benzyl ester are also C1-C15 alkyl esters, or C1-C11 ω-phosphatidylcholinealkyl groups, or three R's are methyl groups and the remaining R's are 05-
Iron represented by ω-phosphatidylcholine alkyl group of C2G? Lufilin derivative complex.

(B) Proximal base type iron porphyrin derivative complex represented by the general formula or methyl group, nFi1-18, m is 1-5) Same, t is 2
2- A proximal base type iron porphyrin derivative complex represented by 10 to 14, n is 1 to 18).

Among the above complexes (A) to (C), the general formula (in which each R is alkyl, ω-hydroxyalkyl, ω-hydroxycarnylalkyl, and an alkyl or benzyl ester thereof having 1 to 5 carbon atoms) A synthetic method is described by J-P.

Co11m1n et al., J, Am, Chem, 5oc
-, 97, 1427 (1975) and meso-tetra-[α, α, α, α-(0-aminophenyl)]porphine, which was synthesized according to the method described in the same document or EPC Patent Application Publication No. 66.884. General formula synthesized according to the publication (however, n is υ as described above, m is a smaller integer, R' is H or 10CH2-(C6H5), R
''￥i alkyl or benzyl group having 1 to 5 carbon atoms)
The acid chloride represented by is reacted at low temperature in the presence of a dehydrochlorination agent such as a tertiary amine or an alkali hydroxide to obtain the corresponding tetraamide. About 5 to 20 times excess FeBr
anaerobically at 90°C in DMF with secondary and tertiary amines.
When heated for about half a day, the corresponding Fs (1'l
l) Porphyrin/Br salt is obtained. R'=-OCH
For 2-(C6H5) or R"=alkyl or benzyl group having 1 to 5 carbon atoms, if excess anhydrous aluminum chloride or alkali hydroxide is applied after introducing iron as described above, the corresponding terminal OH form or terminal -C
Obtain OOH body. However, R'=-OCFI2-(C6H
In the case of 5), better results will be obtained if this operation is performed before introducing iron.

In addition, when R is the above-mentioned alkyl, ω-hydroxylgonylalkyl and its alkyl or benzyl ester, the above-mentioned EPC Patent Application Publication No. 66.8
It is described in more detail in Publication No. 84. In addition, when R is a hydroxyalkyl group, Japanese Patent Application No. 57-210839
Details are given in the specification.

In general formula α), when each R is ω-phosphatidylcholylalkyl, H, J,
Lucas et al. - Chem, & Ind. (Lon
Don) 72.5491 (1950), N, T,
Thuong et al., Bull.

The reaction may be carried out slowly according to the method described in Soc, Chlm, Fr, 1974667, and this method is also applied when R1 to R3 = methyl group and R4 = ω-phosphatidylcholylalkyl group. These iron porphyrin complexes are described in further detail in Japanese Patent Application No. 57-210840 and Japanese Patent Application No. 57-210841.

In the proximal base type iron porphyrin of formula (B), the formula (
The tetraamide type porphyrin (wherein each R is an alkyl group in the formula (~), which is a precursor of the iron porphyrin derivative of A), is treated with an excess of copper (I[) salt, preferably an acetate, in CHCt/CH30H. After reacting below the boiling point to the corresponding copper(n) porphyrin derivative, DMF
-CFTO is introduced into the β-position of virol by 25-reaction with a Vlesmeier reagent such as /POCt5.

This aldehyde is reacted in concentrated sulfuric acid at room temperature for several hours to remove copper, and iron is introduced by the method described above to obtain an iron IO porphyrin derivative. This was reacted with CHCtzCT (NaBH3 in a 3OH mixed solution to obtain the corresponding 1-CH20F1 body, and further phosgene and 1 of the following formula (where t is an integer of 1 to 5 and R to R are hydrogen or methyl groups). A -(ω-aminoalkyl)imidazole derivative is reacted in the presence of a tertiary amine to obtain the desired virol β-position 1 derivative.

In addition, the above-mentioned -CHO form can be converted into the Wittig reagent 2 of the following formula.
After reacting with 6-1, alkaline hydrolysis is performed to obtain pyrrole β.
-CH=CI (-C00T (iron (2) with group
A porphyrin derivative is obtained, this is made into an acid chloride by the action of 5oct2, oxalyl dichloride, etc., and the formula% derivative is reacted with the formula% derivative in the presence of a tertiary amine or an alkali hydroxide to form 1 at the target pyrrole β-position. An iron(t)porphyrin derivative is obtained. Note that the pyrrole β-
If an iron(t)porphyrin derivative having 10H-CI(-COOR at the position of A basic type iron@) porphyrin derivative is obtained at the position. Furthermore, by reacting the above-mentioned 10) I201 (body) with mesityl chloride, -cTr2oso2
Obtain cH3 body, add 10 to 50 times the amount of sodium azide and D
React for several days in MF to obtain -CH2N5 body. This was treated with several times the amount of triphenylphosphine in glyme or dioxane, decomposed with concentrated aqueous ammonia, and -CTI
2NH2 bodies are obtained.

If this is reacted with the imidazole derivative of the following formula using the general Begted condensation method, the desired pyrrole β-position T? 1 Obtain a porphyrin derivative.

The proximal base type Fe(to)porphyrin derivative of formula C) is
(0-aminophenyl)/rufin is reacted and then reacted. The thus obtained porphyrin derivative having an ester group in one of its alkyl chains was introduced with iron using the above-mentioned method, and then subjected to alkaline hydrolysis to convert the ester into -COOH type 1, . By reacting 1-(ω-amino long-chain alkyl)imidazole with a conventional peptide condensing agent, the desired proximal base type Fe(to)porphyrin derivative (C) can be obtained. Note that the ω-amino long-chain alkyl in this case has about 10 to 14 carbon atoms.

In addition, in order for the complex (4) to act as an oxygen adsorbing/desorbing agent, a base needs to be coordinated to the central iron as an axial ligand. (In complexes (B) and (C), such bases are already covalently bonded, so there is no need to add them separately.) Examples of such coordination molecules include the general formula (where Ra, , At least one of R1! is a hydrophobic substituent such as a long chain alkyl group, a trityl group, C1
~C4 alkoxyl? There are hydrophobic imidazole compounds represented by a nyl alkyl group, where ad is hydrogen or a C1-C3 alkyl group. Such an imidazole compound is added in an amount of about 5 to 50 moles or more per mole of iron porphyrin complex.

Example I 5,10,15,20-tetra[α,α,α,α-o-(2Z2'-dimethyl Tetradecanamidophenyl)] Iron forphyllinate (to)
1 mg, 6.5 η of flavin mononucleotide, 83.3 μ of dimyristoylphosphrotidylcholine, and 30-1,34 m 9 of 1-laurylimidazole (axial ligand) were uniformly dissolved in methanol and gently concentrated under reduced pressure. 10 wLt of pure water was added to the obtained film-like material, and the mixture was ultrasonically stirred at 100 W for 30 minutes.

When Na2S2O4 is added to this uniformly dispersed aqueous solution under nitrogen, the visible absorption spectrum changes from 510 nm to 533 nm.
nm, and the reduction reaction was completed in about 30 minutes.

This dispersion was stirred with 5 g of an equivalent mixture of an acidic cation exchange resin and a basic anion exchange resin under nitrogen and filtered.
When a 7.0.0.3 molar phosphate buffer solution, which had been deoxygenated in advance, was added and oxygen gas was introduced, an oxygen complex type spectrum having an extremely thick absorption at 538 nm was immediately obtained. When this material was degassed, oxygen adsorption and desorption was observed with a wide range of 533 nm deoxygenation types. However, when placed under oxygen for 6 hours, approximately 60% was oxidized to FeQIl) type, but when this was subjected to the original reduction operation, extremely thick absorption 53
It returned to the 3 nm Fe(II) type.

■ This aqueous dispersion was developed in an LT (-20 Sephadex column), the distillation curve was determined, and a separately prepared column with a diameter of approximately 20
We confirmed with an electron microscope that it was a liposomal aggregate of OA, and compared it with a distillation curve using an aqueous dispersion of dimylintoyl phosphatidylcholine as a standard.
This sample aqueous dispersion was confirmed to contain liposomes and lamellar aggregates with an average diameter of 250 A.Example 2 5, which was obtained according to the process of Example 24 of EPC Application Publication No. 66.884, 10,15.20-tetra[α,α,α,α-o-(2',2'-dimethyl-
2'-methoxycarbonylacetamido)phenyl]I
Using rufirinato iron Φ00.7■ as an iron surface complex,
The same procedure as in Example 1 was conducted except that 81 ng of nicotinamide adenine dinucleotide was used as the reduction aid. Visible absorption spectrum maximum 510 before adding reducing agent
The wavelength shifted to 534 nm, and the reduction reaction was completed in about 25 minutes.

Thereafter, in the same manner as in Example 1, it was confirmed that the Kutle maximum at 537 nm appeared in the oxygenated visible absorption spectrum.

Note that, similar to Example 1, formation of liposomes and lamellae was confirmed.

Example 3 (A) 100I of 10-dibromodecane and an equivalent amount of sodium benzyl oxide were reacted under reflux in tetrahydrofuran, and the precipitate was filtered, concentrated, and then distilled under reduced pressure to obtain 10-dibromodecane.
Lithium dianion of 2-methylpropionic acid was generated using benzyloxydecanyl bromide diisoprobilamide, and 18 g of the above 10-benzyloxydecanyl bromide was added dropwise at -20°C, followed by reaction at 45°C for 2 hours. Ta. The reaction mixture was added to cold dilute hydrochloric acid, extracted with ether,
The separated ether layer was washed with dilute hydrochloric acid and then with water, separated and dried over Glauber's salt.

The crude oil obtained by evaporation to dryness was recrystallized from petroleum ether to obtain colorless crystals of 12-benzyloxy-2,2-dimethyldodecanoic acid. Melting point 53-55°C. Elemental analysis: C
Calculated value as 21H3403 (@; C75,40, H1
0,25, analysis value (a); C75,64°33-Hlo, 09° proton nuclear magnetic resonance spectrum (cDc
t5) δppm: 1.18 (6H, s*-
CH3).

1.26 (16T(,s, -CH2-), 3.46
(2H2t.

Phenyl-CI (20 mark 2CH2-) y 4.51
(2Hps +7 enil C town 0-), 7.33
(5H,s, phenyl proton).

Got this Cal? 3.34II of phosphoric acid to anhydrous benzene 5
Dissolve ml K and add 1.2 rnl of thionyl chloride,
The reaction was allowed to proceed at room temperature for 12 hours, and the mixture was dried under reduced pressure to obtain 12-benzyloxy-2,2-J methyldodecanoic acid chloride as a colorless oil. Infrared absorption spectrum (CC64) Si1
790 m-'(-'t-ct).

Proton nuclear magnetic resonance spectrum (CDCl3) δpp
m: 1.28 (22H, s, -CH3,- and -〇blood-).

3, 46 (2H, t, PhCH20 (J ridge CH2
-), 4.50 (2) I, s, PhC Town 0
-), 7.32 (51(,s, phenyl proton).

(B) 5,10,15.20-tetra (α, α, α
, α-0-aminophenyl)porphine (hereinafter referred to as H2Ta
(referred to as mPP) 1. OF anhydrous tetrahydrofuran (4
0m/) 34- Make a solution and add 0.81 g of pyridine to 12 obtained in step ① above.
-Benzyloxy-2,2-dimethyldodecanoic acid chloride (3.53.9 g) was added dropwise and reacted for 3 hours. After extraction with ether and washing with water, the separated ether layer was dried with Glauber's salt and dried under reduced pressure. The crude product obtained was purified by silica column chromatography using a mixed solvent of benzene and ether (volume ratio 15:1) to give a brown color. 5,10,15.20-tetra[α,α,α,α-o-(12-benzyloxy-2
゜2-dimethyldodecanamido)phenyl]porphine was obtained. Infrared absorption spectrum (CHCl3) ν3340.

3330 # 3000.2930.2860.168
0°1580.1510.1450.1300.110
0°970.910,700z Proton nuclear magnetic resonance spectrum (coct5) δpp
m knee 2.6 (2H, st porphyrin ring)
-0,23(24H,s, -C(C wholesale2CON)I
-), 3.467, 32 (20H=s)
s 8.82 (8H-m).

(6) 5,10,15.20-tetra[α,α obtained in (B) above.

α,α-o-(12-benzyloxy-2,2-dimethyldodecanamido)phenyl]porphine 1.68.9
25ml of anhydrous dichloromethane and 25ml of nitromethane
After adding Anisole 2 #I/, 2 g of anhydrous aluminum chloride was reacted at the addition mark for 4 hours. Pour into 100 g of ice water to decompose excess aluminum chloride, extract with dichloromethane, wash the separated dichloromethane layer with water, then wash with 10 g of sodium bicarbonate aqueous solution, separate, dry with sodium sulfate, and concentrate under reduced pressure. did. The residue was recrystallized from benzene to give 5. purple plate-like crystals.
10.15.20-tetra [α, α, α, α-o-(1
2-Hydroxy-2,2-dimethyldodecanamido)phenylporphine was obtained in a yield of 80%. Melting point 127-1
29.5℃.

Magnetic field desorption mass 7 (Ktor: 1579 (M+1)" Infrared absorption square (KBr) 3600-3350 (wide), 3440.3330.2940.2860゜16
90. 1585. 1515. 1450. 130
2°1060.970,810. ．． 770.740m-
1° Proton nuclear magnetic resonance spectrum (CDCl3) δ
ppm Ni 2.59 (2H* s t pyrrole; NI
(), -0,22 (24H, s, -C (C town) 2-
CONH-), 3.64 (8H, t, I (QCC
OCH2-), 7.15 (4H-s).

7.36-8.73 (16H', m), 8.8
2 (8H'.

S). Note that 4,50 (8H) derived from a benzyl group.

The absorptions of s) and 7.32 (20H, ti) disappeared.

Elemental analysis: Calculated value (A) as C100H138N+308; C76,00, H8,80, N7.09: Analysis value (4); C75,62, H8,90, N709° (D) Above (6) Te obtained 7' C5, 10, 15, 20
- Tetra [α, α.

α, α-o-(12-hydroxy-2,2-dimethyldodecanamido)phenyl]porphine 0,65I was dissolved in anhydrous tetrahydrofuran (50 m), and pyridine 0.
Ferrous bromide tetrahydrate 2 was added and replaced with nitrogen.
．． OI was added and the mixture was refluxed for 3 hours under nitrogen. After extraction with chloroform and washing with water, the separated chloroform layer was dried with Glauber's salt and the solvent was distilled off under reduced pressure.The resulting residue was subjected to alumina column chromatography using a mixed solvent of chloroform/methanol (volume ratio 9/1). Purified. The eluate was stirred with 2 ml of 48 thihydrobromic acid, dried with Glauber's salt, and evaporated to dryness to give a black-purple solid of bromo(5,10,15,20-tetra[α,
α,α,α-o-(12-hydroxy-12,2-dimethyldodecanamido)phenyl]porphinato)iron(2
) was obtained in a yield of 5't%. Melting point 76-79°C.

Magnetic field detachment mass 7 (Kuttle: 1713 (M+1)" but molecular formula C1oo) 1134N606FeBr =
As 1712.

Infrared absorption spectrum (KBr) 3600-3150
(broad), 3440 m 2930. 2
860, 1690°1580.1slo, 1440.
1300.1075°1000.805,760,71
5cIn.

Elemental analysis: C100T (134N806 ・FsBr
Calculated value (4): C70.13, H8.00,
N6.54, analysis value (to); C70,37,) I g,
40, N 6.63° (g) The same procedure as Example 1 was used except that 0.5 m9 of the iron (2) volf 38-ylin complex obtained in the above ① was used as the metal complex and 7 rng of flavin adenine dinucleotide was used as the reduction aid. A similar operation was performed. The visible absorption spectrum maximum of 510 nm before adding the reducing agent shifted to 534 nm, and the reduction reaction was completed in about 25 minutes ◎ From then on, the same procedure as in Example 1 was carried out until the appearance of the oxygenated type visible absorption spectrum maximum of 537 nm. It was confirmed.

Note that the formation of liposomes and lamellae was confirmed in the same manner as in Example 1.

Example 4 (A) Bromo (5,10,15) obtained in Example 3)
,20-tet,[α,α,α,α-o-(12-hydroxy-2,2-dimethyldodecanamide]phenyl]
Porphinato) iron ([11) 0.15g was dissolved in anhydrous dichloromethane (10m/), and triethylamine 0.0
7ml and 8g of 2-chloro-2-oxo-1,3,2-dioxaphosphorane were added and reacted at room temperature for 12 hours. The reaction was completed by silica dull thin layer chromatography (solvent chloroform/methanol = 1071).
The disappearance of Suit with an Rf value of 0.30 and TLf ((fi,
This was confirmed by the generation of a spot of 0.40. The solvent was distilled off under reduced pressure, the residue was dissolved in 25 ml of anhydrous acetonitrile, and 5 ml of trimethylamine was added at -60 to -40°C, sealed in a pressure-resistant stainless steel container, heated to 60 to 65°C, and reacted for 16 hours. Ta. The black precipitate obtained by cooling to room temperature and filtration was dissolved in methanol and added to Sephadex LH-
The eluted solution was evaporated to dryness and then vacuum-dried in the presence of phosphorus pentoxide. As a black solid,
Iron g) -s, 10+'15+20-tetra
(α, α, α, α −o −C12” (2'
-trimethylamineethyl)phosphoryloxy-2,2-dimethyldodecanamide]phenyl)I
A rufin complex was obtained with a yield of 90%.

Infrared absorption spectrum (KBr) Niji 3650-315
0 (broad), 3430, 2940, 2B
60, 1690.

1580.1510,1440,1220,1080.
1000゜950.800,760,710crt+
.

Visible absorption spectrum (H2O): λmax 412°5
64 nm 0 (B) 0.4 m9 of the complex obtained in (A) above was used as the iron-faced complex, and 210 m of vitamin B was used as the reduction aid.
The procedure was the same as in Example 1 except that 9 was used. The maximum visible absorption spectrum before adding the reducing agent is 512 nm.
33 nm, and the reduction reaction was completed in about 30 minutes.

Thereafter, in the same manner as in Example 1, the appearance of an oxygenated visible absorption spectrum maximum of 537 nm was confirmed.

In addition, as in Example 1, liposomes and La41- Formation of mela was confirmed.

Example 5 (A) 2.1 g of H2TamPP was dissolved in dichloromethane (300 ml), 1 ml of pyridine was added,
Then, the 12-benzyloxy- obtained in Example 3(A)
1.09 p of 2,2-dimethyldodecanoic acid chloride was added dropwise and the mixture was reacted for 3 hours at room temperature, and then 5 ml (large excess) of piparoyl chloride and 5 ml of pyridine were each added dropwise and allowed to react for an additional 2 hours. After that, add saturated aqueous sodium hydrogen carbonate solution (200 ml) and dilute the separated organic layer with 4% hydrogen carbonate.
It was washed twice with Om7 and dried with Glauber's salt. The residue obtained by drying under reduced pressure was dissolved in the solvent tositebenzene/ether = 771
(v/v) was purified by silica dull column chromatography, and further recrystallized from an ether-methanol mixed solvent to obtain 5.1 reddish brown crystals.
0.15.20-()su(α,α,α-〇-biparamidophenyl)-α-o-(12-benzyloxy-2,
2-dimethyldodecanamido)phenyl]42-porphyrin was obtained in a yield of 33%. Melting point 87.5-89
℃. Magnetic field desorption mass spectrum M+1242 (C80H9
ON805=1242), infrared absorption spectrum (
KBr) ν: 3440, 3330, 2960, 294
0°2860.1690,1580.1'510,14
50,1300°1160.970,810,760,
740crn-', proton nuclear magnetic resonance spectrum (
CDC43) δppm: -2,60 (4Hrs
y porphine ring〉N 其), -0,21(6H, s
, -CH2C(CH3)2CONH-) 0.05
, 0.09 (27H, each s + (CI(s)
3C-CONH-), 3.46 (2H, t, J =
6.4 Hz, PhCH20CH2CH2-),
4.50 (2H, s, PhCH20-), 7.8
5 (5H.

S, benzyl group phenyl proton). Elemental analysis: C80
Calculated value (%) as H9ON805: C77,26°I
F (7,30, N9.01, analysis value (chi): C76,
89°) (7,31,N8.88°(B) The product obtained in (4) above, $0.659, was mixed with dichloromethane (15Ttl)-nitromethane (15m
To the mixed solvent solution of (1), 1 ml of anisole and 2 g of anhydrous aluminum chloride were added, and the mixture was allowed to react at room temperature for 3 hours. Pour into ice water (200ml), extract with 50ml of chloroform, and add the separated chloroform layer to water (100ml), 4
% sodium bicarbonate aqueous solution (2 times x 100 ml) and dried with Glauber's salt.

The residue obtained by drying under reduced pressure was dissolved in chloroform/ether =
By purifying by silica dull column chromatography with a 10/1 mixed solvent and further recrystallizing from a benzene-n-hexane mixed solvent, 5.1 of reddish-purple crystals were obtained.
0,15.20-[: )su(α,α,α-0-piparamidophenyl)-α-〇-(12-hyrroxy2,
2-Dimethyldodecanamido)phenyl]porphine was obtained in a yield of 43g1 and 72%. Melting point 220-22
1℃.

Infrared absorption (KBr) = 3650-3200
(broad), 3440, 3330, 2960, 29
30°2860.1690,1580,1510,14
45,1300°1160.970,805,750.
7406n, proton nuclear magnetic resonance spectrum (CDC
tg) δppm Ni 2.60 (2H, s, porphine ring: 17NH), -0,19 (6H, s, -C
H2CH2C ((Jha)2-CONH-), 0.10
, 0.06 (27H, each, (CH3)3
CCON) (-), 3.79 (2H, t, J=6
．． 5Hz, HOCH2CH2-), 8.82 (8H
181 porphyrin ring β-position proton).

Elemental analysis: Calculated value as C7sHa2NaOs (忤)
; C76.01, H7.34, N 9.72, analysis value;
C76,17, H7,46, N 9.43° (C)
0.575 g of porphine obtained in the above (B) was dissolved in 0.37 nl of pyridine and 40 d of tetrahydrofuran,
Under a nitrogen gas atmosphere, 0.6 g of ferrous bromide tetrahydrate was added, and the mixture was reacted under reflux conditions for 4 hours.

The residue obtained by drying under reduced pressure was mixed with chloroform/methanol = 5
Purify by alumina column chromatography using a 0/1 mixed solvent, and add 1 ml of 48 thihydrobromic acid to the elution solution.
was added and stirred, followed by drying with Glauber's salt. After drying under reduced pressure,
The residue was recrystallized from a methanol/dichloromethane mixed solvent to give black-purple crystals of bromo(5,10,15,20-(1
-li(α,α,α-0-biparamidophenyl)-α-o
-(12-Hydroxy-2,2-dimethyldodecanamido)phenyl]porphinato) iron(t) is obtained i0.56
g, 45- was obtained in a yield of 87 g. Melting point 235-237°C. Magnetic field separation max 4 kutor (M+1) 1287°C (M+1)-B
r) 1207 (C7sHa2NaOs, FeB
r=1286, Fe=57.11r=79)
. Infrared absorption spectrum (KBr) ν: 3440,29
70°2940.2B60,1690.1585,15
10,1445°-1- 1300,1260,1000,810,760ctn
, 7C elementary analysis: C73H82N805, Fe Br
Calculated value (ch); C68,11, H6,42, N
8.70, analysis value (chi); C67.84, H6.46
, N 8.58゜(c) 0.5Fl' of the iron(t) complex obtained in (C) above, 37■, 1-lauryl-2 in the vitamin
- Using 1.5 μm of methylimidazole, a homogeneous liposome dispersion was obtained in the same manner as in Example 1, and the reducing agents were 1 m9 of NADP, 10 m9 of glucose-6-phosphate, 0.02 m of ferredoxin, and ferredoxin-NADP.
Reductase 0.1■, catalase 0.05m9 and glucose-6-phosphate dehydrodanase 0.081n
A reduction reaction was carried out using 9. Approximately 60 minutes to the ridge
The visible absorption spectrum maximum of 46-511 nm of type 01D disappears, and the maximum of 531.56 nm of type Fe(II) disappears.
The wavelength has shifted to 3 nm.

After that, in the same manner as in Example 1, however, oxygen was introduced as it was without using an ion exchange resin, and the oxygenated type spectrum was 543.
The appearance of nm was confirmed.

Note that the formation of liposomes and lamellae was confirmed in the same manner as in Example 1.

Example 6 Bromo C5,10°15+20 obtained according to the process of Example 22 of EPC Publication No. 66884
-tetra(α,α,α,α-o-(12'-hydroxycar?nyl-2/, 2/-dimethyldodecaneamide)
The procedure was the same as in Example 3 except that phenyl)porphyrinato)iron ([00,5■) was used.The maximum visible absorption spectrum before adding the reducing agent was 511 nm and 533 nm.
The reduction reaction was completed in about 30 minutes.

Thereafter, in the same manner as in Example 1, the appearance of an oxygenated visible absorption hardness maximum of 536 nm was confirmed.

Note that the formation of liposomes and lamellae was confirmed in the same manner as in Example 1.

Example 7 (A) 20.2 g (20 mmol) of H2TamPP was dissolved in 1.51 g of chloroform and heated under reflux at the boiling point.
u(CH3CO2)z41206. O,! 9 (30
A saturated methanol solution containing 1 mmol) was added. 3
After refluxing for 0 minutes, the mixture was concentrated under reduced pressure, and methanol was added for crystallization. Recrystallization from chloroform-methano-AI gave a copper (rl) complex of H2TamPP.

Yield 20.1 g (yield 93.8%), melting point (mp))
300℃.

TLCIZf=(3,49(silica)fk plate,
Penze.

1690 (νC=O+amide) c
m-1 and other visible spectrum (CHCL3) 2-ax411, 5
34.

568 (shoulder absorption) nm FDMS spectrum m/e 1071 (M+:) Elemental analysis (as C64H64NBO4Cu) Analysis value (calculated value) H: 5.87 (6,01), C near 1.4
0 (71,65), N; 10.29 (10,44
) H. The above copper(n) complex 19. Og (17,7 mmol)
was dissolved in 1.5 l of dichloromethane. Separately DMF68
, 5 ml (0,885 mol) and 4 g POC 82.
Vie 1 sme ie prepared by mixing 5 ml (0,885 mol) under water cooling at room temperature or below
The r complex was added dropwise to the above solution over 30 minutes at room temperature. 8
After boiling under reflux for an hour, the temperature was returned to room temperature. After pouring the obtained dark green immonium salt solution into 1.5 liters of ice water, 500 d of concentrated ammonia water was added at room temperature and reacted for 1 hour. After purification with a silica dull column (7αφX35crn) using a benzene/ether (volume ratio 2/1) solvent, recrystallization from acetone-methanol yields 2-formyl TamPP.
A copper(II) complex was obtained.

Yield 12.8g (yield 65.7'%), mp
260-262°C TLCRf=0.2.6 (Silica dull plate.

Benzene/ether (1/1 (v/v)) IR spectrum (KBr) 1690 (vC=0.amide).

49-1675 (νC=Or aldehyde)tM other visible spectrum (CHCL3) 2maX423, 545.

586 nm FDMS spectrum m/e 11099 (,) Elemental analysis (as C65H64N805Cu1) Actual value (initial value) H: 5.88 (5,86), C near 0.66
(70,92), N: 9.96 (10,18) 2-formi fiv TamPP copper (■) complex 6.
0. ! i+(5,'6 miIJ mole) in 120ml of concentrated sulfuric acid
1 and allowed to react at room temperature for 2 hours. This was poured into 7° 5N aqueous ammonia/dichloromethane (1/1 ratio) under water cooling. The chloroform layer was washed with water, dried and evaporated to dryness under reduced pressure.
5,5crnφX35cr++). Recrystallized from acetone-methanol to give 2-formyl TamPP.
I got it.

Yield 3.54g (yield 60.9%), mp2
58-260°C TLCRf=0.44 (Silica dull grade.

50- Hair -1! ! "n/: I--thyl/acetone (8
/8/1(v/v/v)))IR,(Pect#(KBr
) 1690 (νC=O#amide).

1675 (c-o, aldehyde) crn and other visible spectrum (CHCL3) λmBz 429 r 52
1 +559.599,655nm FDMS spectrum rrv's 1038 (M, )
Elemental analysis (as C65H66N805) Actual value (calculated value) H: 6.16 (6,40), C near 4.83
(75,12), N: 10.65 (10,78)%
PMR spectrum (CDcts) δ (ppm) −2
, 41 (-double line, 2H, pyrrole N-H), 0.068
, 0.120° 0.125 (each - double line, 36H, -
CH5), 7.09-8.87 (multiplet, 26
H, phenyl ring, porphy CMR spectrum # (cDc
ts) δ (ppm) 26.45 (-CH3), 3
8.95, 38.84 (-C(CH3)'x),
115.07-149.94 (phenyl ring, porphyrin ring carbon).

175.39, 175.51, 175.60 (-CON
H-).

18) 1.69 (-CIIO).

(C) 2-formylTamPP 2.299 (2,2
0 mmol) in chloroform/methanol (volume ratio 5/
1), and after adding NaBH4330Wl (8.72 mmol), the mixture was reacted at room temperature for 10 minutes.

Chloroform was added to this, washed with water, dried under reduced pressure, and the residue was recrystallized from chloroform-methanol to obtain sita-hydroxymethyl TamPP.

Yield 2.22g (yield 96.8%), mp) 300℃T
LCnf=o, s 1 (chloroform/methanol (1
0/1 (v/v))) IR Suiktor (KRr) 1690 (v(zQ, amide).

1060(ν(-6H)ctn et al. 1675(shc
-Q H aldehyde)6n Disappearance visible spectrum (C11C13)λrnaX417,5
11°542 (shoulder absorption), 585,640nm FDMS
Spectrum m/s 11040 (,) Elemental analysis (C65
H6sNaOs) Actual value (calculated value) I(: 6.
80 (6,58).

C: 74.78 (74,97), N: 10.65 (
10,76) Chi PMR spectrum (CDCl2) δ(p
pm) -2,62 (- double line, 2H, pyrrole N-H)
, 0.036, 01051゜0.058, 0.0
83 (respectively - doublet #36H, -CH3).

2.79 (triple line, H, -0) (), 4.95 (
Double line.

2H, -CH20H), 7.15-8.83 (multiplet, 26H.

phenyl ring, porphyrin m hydrogen, -co■-).

38.87 (-C(CH,)3), 60.15 (
-CH20H), 114.02-148.20 (phenyl ring, porphyrin i carbon).

175.51, 175.77, 175.90 (-
CONH-).

Visible spectrum (cHcz3) λm, 418,512
°544 (shoulder absorption), 586,643 nm FDMS spectrum m/e 1040 (M+1).

Elemental analysis (as C65H69N904) Actual value (calculated value) H: 6.66 (6,69), C near 4.
87 (75,05), N: 12.04 (12,12)
1% PMR spectrum (CDCts) -2,61(-
Heavy line, 2H.

Pyrrole N-H) -0,083-0,073+0.0
53゜53-0.041,0.036 (each-weight, 36H1-CH
3).

1.92 (wide line, 2H, -NH2), 4.16
(-double line, 2H2-CH2-), 7.19-8.8
2 (multiplet, 27H, phenyl ring, porphyrin ring hydrogen, -CONTr-).

(Parrot 2-formyl TamPP 104m9 (0,
10 mmol) in 20 mJ of dichloromethane and
'CK cooled. After adding 0.23 mmol of carbon tetrachloride solution containing 1.0 mmol of phosdan, the mixture was allowed to react for 1 hour.

After returning to room temperature and reacting for an additional hour, the solvent and excess phosgene were distilled off under reduced pressure and dried to quantitatively obtain 2-chlorocal-nyloxymethyl TamPP in the form of hydrochloride.

TLCRf=0.68 (silica gel plate, benzene/ether/acetone (volume ratio 515/1).

2-formyl TamPP has Rf=0.44FD under the same conditions.
MS m/s 1102 (M, )■) 2-chlorocarbunyloxymethyl TamPP 110 m9
(0.10 mmol) was dissolved in dichloromethane 2 (17), 1-(3-aminopropyl)imidazole 1251
n9 (1 mmol), 54-triethylamine 0.14 mA! (1 mmol) was added and reacted at room temperature for 16 hours. After distilling off the solvent under reduced pressure, it was washed with water, dried, and diluted with chloroform/methanol (10/1 (V
/v)) Silica Dull column using solvent (2crnφX
By purifying with 20cn1), yield i75■ (yield 59.5%) TLCRf=0.39 (silica gel plate, chloroform/methanol (10/1 (v/v)))
IR spectrum (KBr) 1720 (νC=OI urethane), 1690 (νC=Or amide) crn and other visible spectrum (CHCL3) λml! 418, 5
12.

544 (shoulder absorption), 585.640 nm elemental analysis (C7
2H77N1106) Actual value (calculated value) H:
6.37 (6,51), C; 72.23 (72,
52), N: 12.78 (12,92)%P
MR spectrum (CDC23) δ (ppm) -2,6
0 (-double line, 2H, virol N-H), 0.007
, 0.051.

0.075, 0.117 (respectively - double line 136Hj
-CH5).

2.03 (triple line, 2 H, 0CONHCH2CH2
CH2).

3.19 (quartet, 2H,0CONHCH2CH2
CH2), 4.09 (triple line, 2H, 0CON
HCI (2CH2CH2), 5.04 (double line,
H, CH20CO group (), 5.73 (multiplet, 2H9
CH20CONH), 7.03-8.83 (multiplet, 30H, phenyl ring, porphyrin ring, and imidazole ring hydrogen, -CONH-).

CMR spectrum (CDCl2) δ (ppm) 26.
45(-cH3).

31.43, 37.7B, 44.21 (CH2C
H2CH2); 38.89(-C(C)I5)5)
, 61.32 (CH20CONH) , 114
．． 37-151.22 (phenyl ring, porphyrin ring,
and imidazole ring carbon), 156.37 (-0CO■
-).

175.46, 175.72, 176.16 (-
CONH-).

(F) H2TamPP (CH20CONH(CH
2) 3N"'N) 3s ■ Hi-Go (0,032 mmol) was dissolved in 10 ml of THF and heated to FeBr2.4H2092 under boiling point reflux in a nitrogen stream.
, 1rIQ (0.32 mmol) and pyrinone 0.026
ml (0.32 mmol) was added. After reacting at the same temperature for 2.5 hours, it was dried under reduced pressure, and a silica dull column (2
TLCRf=0.27 (Silica gel grade, chloroform/methanol ( 10/1 (v/v) ) )I
R spectrum (KBr) 1725 (νC=Or urethane).

1690 (c-o, amide) crn-1 and other visible spectrum (CHCL3) λm,! 41715051575.
640.657 (shoulder absorption) nm elemental analysis (C72
H75N1106Fe1Br1) Actual measured value (calculated value) H: 5.60 (5,70), C: 64.80
(65,21), N; 11.38 (11,62)%
In addition, this compound is a base-type iron(t)porphyrin complex in which n is 1 in formula (B).

(G) The same procedure as in Example 4 was performed except that the proximal base type Fe(t)porphyrin derivative 0.41n9 obtained in the above procedure) was used. The reduction reaction completed in about 60 minutes, and as a result of introducing oxygen gas in the same manner as in 57-, the absorption maximum reached 548.
The oxygenated sectol of nm was given.

Note that the formation of liposomes and j-mera was confirmed in the same manner as in Example 1.

Example 8 (4) 2-formyl TamPP obtained in Example 7 (A)
55 m9 (o, os mmol) of the M(U) complex were dissolved in 25 ml of toluene, and (Q]P=C)(-CO2CH
After addition of 400 m9 (1.2 mmol) of 5 (mp 162-163 DEG C.), the mixture was refluxed at boiling point for 20 hours. After reaction, 10%-
After washing with an aqueous citric acid solution, water, a 5% aqueous sodium carbonate solution, and water in this order, it was dried over magnesium sulfate, and then dried under reduced pressure. The residue was transferred to a silica dull column (2 columns φ x 20 cm) using benzene/ether (volume ratio 1/1) as a solvent.
) to obtain cis- and trans-integrated Cu(11)-TamPP (CH=CT(-CO
2CH3) was obtained.

(1) trans-Cu(II)-TamPP(CH-C
I (-CO2CT (3) Yield 1%t 54. Om9 (Yield 58.4%) TLCRf = 0.33 (Silica) fk plate.

58- Benzene/ether (Yong Jlz 415).

IR resistance (KBr) 1725 (νC=Or ester).

1690(ν(!xo ramide), 1625(νc=
c.

Visible spectrum (CHCL3) λn1. ．． 422
, 542.

581 nm FDMS spectrum m/e 1156 (M,) Elemental analysis (as C6aH6aNs06Cu1) Actual value (calculated value) H: 5.78 (5,92), C near 0.33
(70,60), N; 9.60 (9,69) Chi (1
1) cis-Cu(lI)-TamPP(CH2CH
2L O2CH3) Yield 9.3■ (Yield 10.1H) TLCRf=0.47 (Silica gel plate, benzene/ether (volume ratio 415)).

IR spectrum (KBr) 1720 (νCwQ1 ester).

1690 (νC=O*amide), 1630 (νC=(+
Visible spectrum (CHCL3) 2m11z 416+5
38+574'nm FDMS spectrum m/e 1156 (M, ) Elemental analysis (as C65H6sNs06cu1) Actual value (calculated value) II: 5.85 (5,92), C near 0.39 (70,60), N: 9 .55 (9,6
9) H (B) Trans-Cu(I) obtained in (A) above
l)-TamPP(CH=CH-CO2CH3) 54
m9 (0,047 mmol) was dissolved in 20 portions of THF. After adding 100 Tv of palladium supported on activated carbon and carrying out a catalytic reduction reaction under a hydrogen stream at room temperature and normal pressure for 6 hours, the catalyst was removed, and then the solvent was distilled off under reduced pressure.

The residue was transferred to a silica gel column (2.8 crnφX4.0 cf.
n), the copper(It) complex of 2-(2-methoxycarbunyl)ethyl TamPP (Cu(Il)
)-TamPP (CH2CH2-CO2CH3) was obtained.

Yield 35.5 Da (yield 65.6%) TLCRf=O150 (silica gel plate, benzene/ether (volume ratio 1/1)).

IR, z, a vector (KBr) 1730 (νC=
= Or nisterurium 1690 (νC = Or amide) crn
Others, 1625 visible spectrum (CHCL3) λm@
z 415 r 538 +572 (shoulder absorption), 622
nm FDMS spectrum m/e 1158 (M,) Elemental analysis (as C68H7oN806Cu1) Actual value (calculated value) H: 5.89 (6,09), C; 70.2
0 (70,48), N: 9.44 (9,67)
%(C) Cu(II)-TamP obtained in (B) above
P(CH2CH2CO2CH3) is alkaline hydrolyzed and returned to acidic Cu(Ir)-TamPP(CH2CH2
COOH) was obtained. 15m9 (0,013 mmol) of this was dissolved in 10 parts of dichloromethane to give 0.0% of thionyl chloride.
0.5m (0.69 mmol) was added. After boiling under reflux for 2 hours, the mixture was dried under reduced pressure.

Redissolve the residue in dichloromethane lQm and cool on ice at 0-5°C.
A solution of On the other hand, 1-(5-aminopentyl)-2
-Methylimidazole dihydrochloride (mp 142-1
43°C) 3.21n9 (0,013 mmol) was dehydrochlorinated according to a conventional method and dissolved in 5 ml of dichloromethane containing 0.01 ml (0,C172 ml) of triethyl allo-1-mine. This was added dropwise to the above solution and reacted at 0 to 5°C for 1 hour, then returned to room temperature and left overnight. After washing twice with the same amount of water, it was dried and concentrated under reduced pressure. The residue was dissolved in chloroform/methanol (10/1 (v/v)
) ') Silica gel column using solvent (2cfnφX
20 cm).

Revenue! 12.0Tn9 (70.8%) TLCRf=0.27 (Silica Dull Plate, Chloroform/Methanol (Volume Ratio 10/1)) IR Suictor (KBr) 1690.1665 (νC=O*amide)
m″″1 and others, 1710 (νc=Q l carnic acid) ff
i””disappeared.

Visible spectrum (CHCL3) Good luck! 415, 53
7.

572 (shoulder absorption) y620hm Elemental analysis (as C76H83N1105Cu1) Actual value (calculated value) T(: 6.21 (6,46).

62-C: 70.28 (70,54), N; 11.6
9 (11,90)%) CH3 (0,007 mm! J mole) obtained in the above (C) was decoppered and ionized in exactly the same manner as in Example 7 (B), and then When iron ions were introduced according to (F), an iron valence complex was obtained in exactly the same manner.

Yield: 7.1 m9 (Yield: 74.2 cm) TLCRf=0.24 (Silical Rugrate, Chloroform/Methanol (1o/i (v/v))) IR
Spectrum (KBr) 1690.1665 (νC=0°amide) cnl Other visible spectrum (CHCL!,) 2max 416.
508.

576.640,674 (shoulder absorption) nm.

FDMS mle 1285 ((M+1),
) Elemental analysis (c76Hs; as N11o5F'ejBr1) Actual value (calculated value) H: 5.97 (6,12)
, C: 66.54 (66,81), N: 11
．． 02(11,28)% (E) amide type proximal base FeωD porphyrin derivative obtained in (D) above 0.6 m
The procedure was the same as in Example 4 except that 9 was used. The reduction reaction was completed in about 45 minutes, and the maximum visible absorption spectrum reached 510 nm.
It was confirmed that m shifted to 535,562 nm and became Fe (II) type.

When oxygen gas was introduced into this, it became an acidic type with a maximum wavelength of 544 nm.

Note that formation of liposomes and lamellae was confirmed in the same manner as in Example 1.

Example 9 (n 5,10.15-
Cal xyethyl-2-methyl)propanamido]phenyl)porphyrin 2921n9 (0,273 mmol) in DMv/dichloromethane (volume ratio 1/1) 4omz
Triethylamine is dissolved in 0.2+++ A! at 0°C. (
1,4 mmol), 1-(12-aminododecanyl)imidazole dihydrochloride 103 m9C0,32 mmol), and diethyl phosphate cyanide 62.3 m9 (0,
38 mmol) was added. After 1 hour, the mixture was returned to room temperature and reacted overnight.

After conventional treatment, chloroform/methanol (15/1 (v/
v) ) Silica Dull column using solvent (3 rust φ x 3
8crn) to produce 5,10.15-tri[α,α,α-(0-bipalamide)phenylcou20
-mono(-α-Co-(2-(N-[12-(1-imidazolyl)dodecanyl]carnomoylethyl-2-methyl)propanamide]phenyl)porphyrin was obtained.

Yield 310 m9 (87, 1 section) TLC: Rf=0.54 (Silica Dull plate, chloroform/methanol (volume ratio 15/1)) IR spectrum (KBr) 2930.2860 (νCH2) cm
-1+ 1690 (νC=Or amide), 1660 (”
C=Ol amide) crn-' et al., 1720 (νC=0.
Carganic acid) crn disappeared.

FDMS spectrum m/e 1302 ((M+1)
, ) PMR spectrum (cDct3) δ (ppm
): 2.63 (--65=multiplet, 2H, virol NH), -0.1 to 0.3 (multiplet.

33H, -CH5), 0.6 to 1.4 (multiplet, 24
Hl -CH2-) +3.00 (multiplet, 2H, -
CH2-), 3.94 (multiplet.

2H1-CH2-) + 5.69 (multiplet, I(
, -CONH-).

6.9 to 8.9 (multiple line, 31H, phenyl ring hydrogen, pyrrole β-position hydrogen, imidazole ring hydrogen, -CONH- elemental analysis (as C81H95N1105) actual value (calculated value) H: 7.33 (7, 35), C Near 4.
39 (74,68), N: 11.57 (11,83
)% visible spectrum (C) (C45)λmax417,
512゜544.585,640nm Porphyrin 90 m9 (0.07 mmol) thus obtained was dissolved in 20 ml of THF, and pyridine Q,
1mA! After adding (1,2 mmol), ferrous bromide tetrahydrate 801119 (0,28
mmol) was added and the mixture was refluxed at boiling point for 3 hours. After drying under reduced pressure, the iron trivalent complex (counter ion Br) was obtained by purifying with a silica dull column (2zφX 20 cm) using a chloroform/methanol (volume ratio 12/1) solvent.

66- Yield t601n9 (60.5% ”) TLCRf=0.53 (Silica e#7'l/-)
, chloroform/methanol (volume ratio 15/1)) 11
t spectrum (KBr) 2930.2860 (νCH2
) +1690 (νC=O+ amide), 1660
(νC=O*amide) cIn-1 and others.

FDMS spectrum m/e 1355 (M,) Elemental analysis (as C81H93N1105Fe1Br1) Actual value (calculated value) H: 6.41 (6,53), C;
67.49 (67,73), N: 10.63 (10
,73) CH visible spectrum (CHCL3) λma! 41
6,507°580.642.675 (shoulder absorption) nr
n.

The complex with formula (C) is one in which RR is hydrogen, n is 1, and t is 12.

operated on. Visible absorption spectrum maximum 510 nm is 5
It shifted to the 35 nm Fe(If) type, and the reduction reaction was completed in about 45 minutes. When oxygen gas is introduced into this, 547n
It became the oxygenated form of m.

Note that the formation of liposomes and lamellae was confirmed in the same manner as in Example 1.

Example 10 R, G, In@keep + J, Inorg,
Nuel, Chem, +24763 (1962)
Bis(1,10-phenanthroline) synthesized according to
Copper (In・(PF6)211n9 with duroquinone 5 m
9 and 100 rv of egg yolk lecithin were mixed and dissolved in methanol, and the mixture was gently concentrated under reduced pressure to obtain a film-like substance. A phosphoric acid aqueous solution IQm/ of pH 7.0.15 mol was added to this, and the mixture was cooled to a temperature of 30° C. or lower and then ultrasonically stirred at 60 W for 30 minutes to obtain a uniform dispersion of liposomes.

This material exhibits a blue color and has maximum absorption at about 600 nm. When 1 ta of Na2S2O4 was added under anaerobic conditions, approx.
Completion of the color tone change was observed in 0 minutes, and it was confirmed that the color change had changed to the corresponding copper(I) salt.

Comparative Example 1 The same operation as in Example 10 was carried out except that duroquinone was not used. No change in color tone was observed even after one hour had passed after addition of Na2S2O4, indicating that no reduction had taken place.

Examples 11 to 17 A 1% by weight liposome homogeneous dispersion was obtained in the same manner as in Example 1, except that the metal complex, reducing aid, and phospholipid shown in Table 1 below were used at a molar ratio of 1/201500. Ta. A reducing agent shown in Table 1 was added to this under anaerobic conditions, and the time for completion of reduction was determined by tracking changes in the visible absorption spectrum of the metal complex. The results are also listed in Table 1.

69-

Claims (9)

[Claims] (1) In a method of reducing a fat-soluble metal complex, which is mainly composed of phospholipids and is embedded in the hydrophobic region of the Isome or lamella, in an aqueous medium by the action of an external reducing force, A fat-soluble reducing agent having a negative redox potential than that of the central metal and having reversible redox ability is embedded in the liposome or lamella in advance, and the reducing agent is The central metal of the metal complex having the central metal in an oxidized state is reduced by applying a reducing force corresponding to a negative redox potential to the liposome or lamella from the outside. Method for reducing metal complexes. (2) The reduction method according to claim 1, wherein the central metal in the metal complex has a more positive redox potential than -0.35 zalto. (3) The reduction method according to claim 2, wherein the reduction aid is a quinone compound. (4) The reducing aid is a benzoquinone compound represented by the formula (wherein R1 to R4 are hydrogen, halodane, a pencil group, or an alkyl group, alkenyl group, or alkynyl group having 5 or less carbon atoms, respectively). The reduction method described in Scope No. 3. (5) A patent claim in which the reducing aid is a naphthoquinone compound represented by the formula (wherein R1 to R6 are each hydrogen, halodane, benzyl group, or an alkyl group, alkenyl group, or alkynyl group having 5 or less carbon atoms) The reduction method described in item 3. (6) The reducing aid is an anthraquinone compound represented by the formula (where R1 to R8 are each hydrogen, halogen, benzyl group, or an alkyl group, alkenyl group, or alkynyl group having 5 or less carbon atoms). The reduction method described in Scope No. 3. (7) The reduction aid has the formula (where n-'fx and R12 are each hydrogen).
Rodashi, benzyl group, alkyl group with 5 or less carbon atoms 1, alkenyl group or alkynyl group, or alkyloxycarboxylic group with 5 or less carbon atoms is nyl group, alkylamido group, alkenyloxycarbunyl group, alkenylamide group, alkynyloxy Cal? 4. The reduction method according to claim 3, wherein the merocyanine dye compound is a merocyanine dye compound represented by a nyl group or an alkynylamide group. (8) The reduction aid has the formula (where R to R are each hydrogen, halodane, benzyl group, and each is an alkyl group, alkenyl group, or alkynyl group having 5 or less carbon atoms, and X is oxygen or sulfur)
The reduction method according to claim 3, which is a galocyanine dye compound represented by: (9) A patent claim in which the reducing aid is a flavin dye compound represented by the formula (wherein R1 to R6 are hydrogen, halogen, pendyl group, or an alkyl group, alkenyl group, or alkynyl group having 5 or less carbon atoms). - Reduction method described in Scope 3 - A patent in which the R1 reduction aid is 1 for vitamins, 2 for vitamins, 3.7 for vitamins Ravin mononucleotide, flavin adenine dinucleotide, nicotinamide adenine dinucleotide, hypoxane or taha vitamin B2 Reduction method described in claim 2 @